Regulation Of MiR-21-5p Targeting SCML2 On Proliferation And Aerobic Glycolysis Of Hepatocellular Carcinoma Cells

Research background and purpose:Hepatocellular carcinoma(HCC)is one of the most common malignant tumors in the world,with an increasing incidence rate,especially in China.With the rapid development of surgery,radiotherapy,chemotherapy,radiofrequency ablation,targeted therapy and immunotherapy,the five-year survival period of patients with liver cancer has increased,but the prognosis is still poor,and the five-year recurrence rate is also high.Therefore,it is urgent to understand the molecular mechanism of HCC progression in order to predict and control the disease as soon as possible.Compared with normal differentiated cells that rely on oxidative phosphorylation of mitochondria to produce energy,cancer cells have evolved to use aerobic glycolysis(also known as the Warburg effect)for growth and reproduction,while providing raw materials for other synthetic metabolism.Micro RNAs(mi Rs)can target multiple genes by regulating the translation or degradation of m RNA,participate in signal transduction pathways and other functions that affect the physiological functions of normal or tumor cells,and are related to aerobic glycolysis.The epigenetic changes of mi Rs and the abnormal expression of their target genes may provide potential tools and opportunities for the detection and treatment of liver cancer.Micro RNA-21-5p(mi R-21-5p)is a carcinogenic mi RNA,which is highly expressed in HCC and low expressed in normal liver tissue.It can regulate a variety of biological processes,including cell metabolism.Overexpression of mi R-21-5p has been proved to promote the proliferation and metastasis of cancer cells,but the regulatory mechanism of its proliferation and aerobic glycolysis in HCC is still unclear.This experiment explored the effect of mi R-21-5p on the malignant behavior of hepatocellular carcinoma cells in HCC and the regulation of glycolysis,evaluated the targeted relationship between mi R-21-5p and SCML2 in hepatocellular carcinoma cells,and provided a new theoretical basis for finding the targeted treatment of hepatocellular carcinoma.Methods:1.Through reading the relevant literature and using bioinformatics tools to analyze and screen the up-regulated mi RNAs in liver cancer tissue,select the mi RNAs that express significantly different in normal liver tissue and liver cancer tissue,and define the research object as mi R-21-5p.2.Western blot,q RT-PCR and immunohistochemistry were used to investigate the expression of mi R-21-5p and target gene SCML2 in hepatoma cells.3.Hep G2 cell model was obtained by transfection,and the effect of mi R-21-5p on the proliferation of hepatoma cells was detected by MTT method.4.Use target gene prediction software to predict the possible target genes of mi R-21-5p,and verify the interaction between mi R-21-5p and target genes through luciferase reporter gene experiment.Result:1.MTT colorimetric analysis showed that the high expression group of mi R-21-5p could promote the proliferation of hepatoma cells.2.Western blot,immunohistochemistry and q RT-PCR analysis showed that the expression of SCML2 was positively correlated with mi R-21-5p in hepatocellular carcinoma.3.The mi R-21-5p/SCML2 signal axis controls the progression of hepatocellular carcinoma through aerobic glycolysis demodulation:1)The target gene of mi R-21-5p was selected by using the target gene prediction tool because of SCML2.The online analysis website of Shengxin suggested that SCML2 was highly expressed in HCC and was closely related to the poor prognosis of HCC.2)The double luciferase reporter gene experiment confirmed that mi R-21-5p directly targeted the 3 ′ UTR untranslated region(3 ′ UTR site)of SCML2 and promoted the mRNA and protein expression level of SCML2.In addition,mi R-21-5p interacts with the wild-type 3 ′ UTR site of SCML2,but has no effect on the mutant 3 ′ UTR site.3)Glucose uptake and lactic acid production experiments confirmed that mi R-21-5p promoted the aerobic glycolysis process of hepatocellular carcinoma.4.The expression of SCML2 in tumor tissues and cells of HCC patients increased and was positively correlated with the expression of mi R-21-5p.Conclusion: The expression of mi R-21-5p is up-regulated in HCC,and mi R-21-5p enhances the aerobic glycolysis of HCC cells and promotes the proliferation of HCC cells by targeting SCML2.